Method for controlling gram positive bacteria with dichlorophenylosazones



United States Patent 3,337,397 METHOD FOR CONTROLLING GRAM POSITIVE BACTERIA WITH DICHLOROPHENYLOSAZONES Marvin L. Oftedahl, Crestwood, Mo., assignor to Monsanto Company, St. Louis, M0., a corporation of Delaware No Drawing. Filed Feb. 9, 1966, Ser. No. 526,070 5 Claims. (Cl. 167-30) This invention relates to the control of bacterial growth, and more particularly to the use of certain carbohydrate osazones for such a purpose.

The compounds of this invention having the following formula where n is an integer from 0 to 4. More specifically, the compounds of this invention include triose 3,4-dichlorophenylosazone (where n=0), and the d and 1 forms of the 3,4-dichloropl1enylosazones wherein the asymetric carbon atoms have any one of the glycero, erythro, threo, arabino, lyxo, ribo, xyl-o, altro, galacto, gluco or manno configurations. Particular exemplary compounds are such as d or l-th-reo-pentose 3,4-dichlorophenylosazone, d or l-lyxo-hexose 3,4-dichlorophenylosazone and d or l-glucoheptose 3,4-dichlorophenylosazone.

This class of compounds can be prepared by the well known reaction of a hydrazine with a reducing monosaccharide to produce a sugar osazone.

The following are given as examples of this reaction for the purpose of illustration only and are not to be construed as limiting the scope of the present invention in any manner.

Example 1 A suitable reaction vessel is charged with a solution containing 3.6 grarns (0.02 mol) of anhydrous d-glucose, 12 grams of sodium acetate, 12.8 grams (0.06 mol) of 3,4-dichlorophenylhydrazine hydrochloride, ml. acetic acid, and 10 ml. saturated aqueous sodium bisulfite in 100 ml. 15% (by volume) ethanol. The solution is heated on a steam bath for minutes, removed from the heat and evaporated to half its original volume under vacuum, then cooled. The yellow solid which crystallizes is collected and recrystallized from 35% aqueous ethanol, The product is d arabin-o hexose 3,4 dichlorophenylosazone, the reaction being I CH=NNHOL 01 I g edema-Q01 Qi-QNIn 3,337,397 Patented Aug. 22, 1967 Analysis.-Calculated for c rt cnmop 01, 28.6. Found: Cl, 28.5.

Example 2 Following the detailed procedure set forth in Example 1, the reactants are 3.0 grams (0.02 mol) of d-xylose and 12.8 grams of 3,4-dichlorophenylhydrazine hydrochloride. The product obtained is d-threo-pentose 3,4-dichlorophenylosazone.

As pointed out above, the compounds of this invention have been found to possess useful and unexpected microbiological activity. In this regard, such compounds have been found to be particularly effective in the control of gram positive bacteria. To demonstrate the aforesaid control, the following test procedure was employed. Stock solutions were prepared by dissolving mg. of the compound to be treated in 10 ml. of acetone, alcohol or other solvents. These stock solutions were serially diluted by pipetting 2 ml. of the stock solution into 18 ml. of sterile nutrient agar to obtain a 1x10 dilution and continuing this procedure in the same manner to obtain further dilutions up to 1x10. The agar was poured into Petri dishes, allowed to harden, and was then spot inoculated with one drop of a cell suspension of Staphylococcus aureus. The suspension was prepared by suspending the growth from a 24 hour nutrient agar slant culture in 10 ml. of distilled water. The inoculated samples were incubated at 37 C. for 48 hours and were thereafter examined to determine whether or not bacterial growth had occurred.

When the above test was conducted with representative compounds of this invention, it 'was' found that d-threopentose 3,4-dichlorophenylosazone showed no growth of bacteria at a dilution of at least one part per one hundred thousand parts of diluent; d-arabino-hexose 3,4-dichloro phenylosazone showed no growth of bacteria at a dilution of at least one part per million parts of diluent. Similar results are obtained with other compounds of this invention.

While the invention has been described herein by means of certain specific examples, it is not so limited. It is to 'be understood that variations and modifications thereof may be made by those skilled in the art without departing from the spirit and scope of the invention.

The embodiments of the invention in which an exclusive property or privilege is claimed are defined as follows:

1. A method for controlling the growth of gram positive bacteria by contacting said bacteria with an effective amount of a compound of the formula where n is an integer from 0 to 4.

2. A method as defined in claim 1 wherein n is 2.

3. A method as defined in claim 1 wherein n is 3.

4. A method as defined in claim 1 wherein the compound is d-arabino-hexose 3,4-dichlo-rophenylosazone.

5. A method as defined in claim 1 wherein the compound is d-threo-pentose 3,4-dichlorophenylosazone.

References Cited Chemical Abstracts 59: 11901e (1963). ALBERT T. MEYERS, Primary Examiner. SAM ROSEN, Examiner.

JEROME D. GOLDBERG, Assistant Examiner. 

1. A METHOD FOR CONTROLLING THE GROWTH OF GRAM POSITIVE BACTIERIA BY CONTACTING SAID BACTERIA WITH AN EFFECTIVE AMOUNT OF A COMPOUND OF THE FORMULA 